KDA Awards Five Research Grants

Five research grants were awarded totaling $199,000. Award recipients and summaries are listed below.

Award $50,000
Targeting AR toxicity in SBMA by modulation of USP7 activity.
Anna Pluciennik, Ph.D.
Department of Biochemistry and Molecular Biology
Thomas Jefferson University

Summary: PolyQ-expanded androgen receptor-dependent cellular toxicity in the neuromuscular system is a characteristic feature of SBMA pathogenesis, although the molecular mechanisms for these effects are poorly understood. However, because cellular toxicity in SBMA is likely to arise, at least in part, from the polyQ-expanded-dependent dysregulation of protein-protein interactions that sustain normal cellular function, we reasoned that identification of such dysregulated interactions might help reveal potential therapeutic targets for disease modification. Therefore, as part of a KDA-funded project (2014), we used a quantitative proteomics approach and identified a deubiquitinating enzyme, USP7, that preferentially interacts with polyQ-expanded AR and contributes to toxicity. In fact, we have demonstrated that not only does partial knockdown of USP7 protein expression decrease mutant AR-dependent aggregation and dihydrotestosterone-dependent cytotoxicity, but also that overexpression of the protein aggravates these cellular effects. These results establish the need for further investigation into the role of the deubiquitinating function of USP7 in SBMA pathogenesis. The objective of this -research is to investigate the functional consequence of pharmacological inhibition of USP7 deubiquitinase activity in cellular and mouse models of SBMA. These studies will extend our previous KDA-funded proteomics work and, if successful, will establish the validity of inhibition of USP7 activity as a therapeutic approach for the treatment of SBMA.

Award $50,000
Identification of New Polyglutamine-Specific Mutant AR-Interacting Proteins in SBMA Motor Neurons
Xia Feng
Postdoctoral Fellow, Neurogenetics Branch
National Institute of Neurological Disorders and Stroke, National Institute of Health

Summary: Spinal and bulbar muscular atrophy (SBMA) is a slowly progressive neuromuscular disease. As the disease proceeds, nerve cells in the spinal cord (called motor neurons) start to die and muscle cells will waste away. The causative mutation of SBMA is the mutant androgen receptor (AR) with an abnormal expansion in the certain region. Such aberrant expansion in the mutant protein (called a polyglutamine tract) damages the normal function of the protein as well as obtains toxicity. Thus, it is important to understand how the mutant AR is regulated via other proteins in the disease. Here, I propose to use a human induced pluripotent stem cell model to generate disease-relevant motor neuron-like cells, and use these cells to identify specific interacting proteins of the mutant androgen receptor. I am hoping that my research will help provide a motor neuron-specific basis for designing and developing novel therapeutics for the treatment of SBMA.

Award $33,000
Transcellular regulation of the proteostasis network in Kennedy’s disease
Laura Bott, Ph.D.
Postdoctoral Fellow
Molecular Biosciences, Northwestern University

Summary: Misfolding and abnormal accumulation of the mutant androgen receptor in Kennedy’s disease indicates that the cellular machinery controlling protein abundance, folding, and transport (proteostasis) may be defective in the disease. We are planning to study molecular mechanisms of Kennedy’s disease in the worm Caenorhabditis elegans. For this, we will genetically engineer worms to express the mutant androgen receptor in the neuromuscular system and assess effects of the polyglutamine expansion on proteostasis regulation in this model organism. Insights into cell type-specific effects and regulation across tissues may lead to new therapeutic approaches for this disease.

Award $33,000
One gene, many proteins: investigating the role of AR isoform 2 as a therapeutic target for SBMA
Carlo Rinaldi, MD, PhD
Department of Physiology, Anatomy and Genetics
University of Oxford

Summary: The activity of steroid hormone receptors, such as progesterone and oestrogen receptors, is modulated by a number of isoforms and splice variants in a tissue-specific manner, in both health and disease. Androgen receptor may be no exception. AR isoform 2, or AR45 by the molecular weight of its encoded widely-expressed protein, is the only naturally occurring AR isoform, arising from use of an alternative transcriptional start site in intron 1 of the AR gene and containing a short, unique seven amino-acid-long N-terminal stretch instead of the long N-terminal domain found in the full length AR. Overarching aim of this proposal is to unravel the contribution of AR isoform 2 to SBMA pathogenesis and provide a novel therapeutic target for this disease, suitable for oligonucleotide antisense targeting without running the risks associated with silencing of the only available copy of the AR gene in males.

Award $33,000
The use of induced stem cells and microfluidics for developing new assays to identify new therapies for Kennedy’s disease
Thomas M Durcan, Ph.D., Assistant Professor,
Montreal Neurological Institute, McGill University

Summary: It is important to explore new avenues in the search for treatments for Kennedy’s disease (KD). Our overall goal is to develop disease-relevant assays that use human motor neurons made from stem cells of both men with Kennedy’s disease and healthy individuals. Using special silicone microfluidic devices to grow motor neurons, we will measure the function of neurons upon exposure to different compounds. First, we will develop assays to measure the survival and growth of the motor neurons. Next, we will look at the how well mitochondria work and move within the neurons. Finally, we aim to set up an assay to test the ability of the normal and KD motor neurons to form synapses (junctions). Taken together, these innovative assays will provide a foundation to build a KD drug discovery platform to screen for promising compounds and targets to treat KD.

Is ASO a potential treatment for Kennedy’s Disease?

This is a follow up to my November 15, 2017, article, “MDAAnnounces SBMA Research Grant.” The research paper was a little over my head (nothing new for me), so I asked the KDA’s resident biology professor, Ed Meyertholen, to explain what Dr. Lieberman’s research was about. Below is Ed’s summary of the grant. For a short primer, I have included the link to a video on DNA-RNA.

"The grant the Andy Lieberman received was to continue the research on the use of Anti-Sense Oligonucleotides (ASO) as a treatment for Kennedy’s Disease (KD). To best understand how it works, it is important to remember the following:

1. KD is believed to be the result of a misfolded protein, specifically, the protein known as the Androgen Receptor (AR).

2. Proteins are built of specific sequences of amino acids, thus to make a protein, one must have amino acids and the sequence of the amino acids of the protein of interest.

3. The sequence of amino acids for any protein are hard coded into our genes - our DNA.  Thus to make a particular protein, the cell must find the gene that codes for the sequence for that protein and read the code to get the sequence.  The structure of the cell that makes the protein is the ribosome.

4. In KD, the misfolded protein is known as the Androgen Receptor (AR) and it misfolds because our DNA has an error in the sequence.  So, when our cells want to synthesize the AR, our instructions are faulty and when we make the resulting protein, it somehow causes cells to die albeit, slowly.

5. Protein synthesis requires two major steps, the first is the synthesis of an RNA copy (RNA is like DNA) of the gene (DNA) which codes for the protein of interest (this occurs in the nucleus).  The RNA synthesis is known as transcription.

6. The RNA copy (which contains the code for the protein) leaves the nucleus and goes to the ribosome.  Here the code is read and the protein is synthesized.  This actual making of the protein is known as translation.

7.  An ASO is a specially designed fragment of RNA that binds only to a specific RNA.  An ASO can be designed to bind specifically to any given RNA.  In this case, the ASO binds only to the RNA that is used to make the AR.  When the ASO binds to the RNA, the cell responds by destroying the RNA (that is what it does) - thus the RNA to make the AR is destroyed before the protein is made and thus no AR is synthesized and thus, it is hoped, no KD.

8.  Andy's grant is, as I understand it, will try to test this procedure on mice models of KD and involve investigating the best ways to deliver the ASO.  Let me also add, there have been several published studies that have shown that ASO's are effective in preventing KD in mice.  Other ASO's have been developed to treat other diseases and just recently, one was approved for use in a disease called Spinal Muscular Atrophy (this is not KD)."  

Nonalcoholic fatty liver disease in spinal and bulbar muscular atrophy

Nonalcoholic fatty liver disease in spinal and bulbar muscular atrophy

Robert D. Guber, BS*, Varun Takyar, MD*, Angela Kokkinis, BSN, RN, Derrick A. Fox, MD, Hawwa Alao, MD, Ilona Kats, BA, Dara Bakar, BA, Alan T. Remaley, MD, PhD, Stephen M. Hewitt, MD, PhD, David E. Kleiner, MD, PhD, Chia-Ying Liu, PhD, Colleen Hadigan, MD, Kenneth H. Fischbeck, MD, Yaron Rotman, MD and Christopher Grunseich, MD
Correspondence to Dr. Grunseich: christopher.grunseich{at}nih.gov

ABSTRACT

Objective: To determine the prevalence and features of fatty liver disease in spinal and bulbar muscular atrophy (SBMA).

Methods: Two groups of participants with SBMA were evaluated. In the first group, 22 participants with SBMA underwent laboratory analysis and liver imaging. In the second group, 14 participants with SBMA were compared to 13 female carriers and 23 controls. Liver biopsies were done in 4 participants with SBMA.

Results: Evidence of fatty liver disease was detected by magnetic resonance spectroscopy in all participants with SBMA in the first group, with an average dome intrahepatic triacylglycerol of 27% (range 6%–66%, ref ≤5.5%). Liver dome magnetic resonance spectroscopy measurements were significantly increased in participants with SBMA in the second group relative to age- and sex-matched controls, with average disease and male control measurements of 17% and 3%, respectively. Liver biopsies were consistent with simple steatosis in 2 participants and nonalcoholic steatohepatitis in 2 others.

Conclusions: We observed evidence of nonalcoholic liver disease in nearly all of the participants with SBMA evaluated. These observations expand the phenotypic spectrum of the disease and provide a potential biomarker that can be monitored in future studies.

Photo:  https://www.quora.com/What-is-liver-failure

US scientists try 1st gene editing in the body

In recent years I have reported several times about CRISPR and gene editing. I am not a scientist, but, to a layman, this sounds like the best possibility for curing Kennedy's Disease. In the article referenced below, it discusses the potential benefits and possible dangers of gene editing.

There is much more research that needs to be done, but the possibility of a cure is now closer than ever before.

US scientists try 1st gene editing in the body

"...This time, the gene tinkering is happening in a precise way inside the body. It’s like sending a mini surgeon along to place the new gene in exactly the right location.

“We cut your DNA, open it up, insert a gene, stitch it back up. Invisible mending,” said Dr. Sandy Macrae, president of Sangamo Therapeutics, the California company testing this for two metabolic diseases and hemophilia. “It becomes part of your DNA and is there for the rest of your life.”

That also means there’s no going back, no way to erase any mistakes the editing might cause.

“You’re really toying with Mother Nature” and the risks can’t be fully known, but the studies should move forward because these are incurable diseases, said one independent expert, Dr. Eric Topol of the Scripps Translational Science Institute in San Diego.

Protections are in place to help ensure safety, and animal tests were very encouraging, said Dr. Howard Kaufman, a Boston scientist on the National Institutes of Health panel that approved the studies.

He said gene editing’s promise is too great to ignore. “So far there’s been no evidence that this is going to be dangerous,” he said. “Now is not the time to get scared.”..."

A short APNews animation video does a good job of explaining the process. 

(AP Animation/Marshall Ritzel)

MDA Announces SBMA Research Grant

This week the MDA announced the awarding of more research grants. Of particular interest to those of us living with Kennedy’s Disease (SBMA) is the following grant.

Testing a potential therapy for spinal-bulbarmuscular atrophy (SBMA)

“Scientists at the University of Michigan Medical School in Ann Arbor are completing preclinical studies in a mouse model to establish the safety and efficacy of a new type of therapy to silence activity of the gene that is mutated in SBMA.”

Below is the abstract of the research noted in the above announcement. It should be noted that Dr. Lieberman serves on the Kennedy’s Disease Association’s Scientific Review Board.

Rescue of MetabolicAlterations in AR113Q Skeletal Muscle by Peripheral Androgen Receptor GeneSilencing

Giorgetti E1, Yu Z1, Chua JP1, Shimamura R1, Zhao L2, Zhu F3, Venneti S1, Pennuto M4, Guan Y3, Hung G5, Lieberman AP6.

Abstract

“Spinal and bulbar muscular atrophy (SBMA), a progressive degenerative disorder, is caused by a CAG/glutamine expansion in the androgen receptor (polyQ AR). Recent studies demonstrate that skeletal muscle is an important site of toxicity that contributes to the SBMA phenotype. Here, we sought to identify critical pathways altered in muscle that underlie disease manifestations in AR113Q mice. This led to the unanticipated identification of gene expression changes affecting regulators of carbohydrate metabolism, similar to those triggered by denervation. AR113Q muscle exhibits diminished glycolysis, altered mitochondria, and an impaired response to exercise. Strikingly, the expression of genes regulating muscle energy metabolism is rescued following peripheral polyQ AR gene silencing by antisense oligonucleotides (ASO), a therapeutic strategy that alleviates disease. Our data establish the occurrence of a metabolic imbalance in SBMA muscle triggered by peripheral expression of the polyQ AR and indicate that alterations in energy utilization contribute to non-neuronal disease manifestations.”

Life Doesn’t End With The Diagnosis

I remember when I was diagnosed with ALS. I could not have been more devastated. Then, a few years later, when my diagnosis was corrected to Spinal Bulbar Muscular Atrophy, it was initially a relief. It did not take long, however, for reality to settle in. I had Kennedy’s Disease!

My symptoms appeared in the late 20s. I am now 70. When I look back at those first days after finding out I had KD, I have to laugh. Life did not end with the diagnosis. It began.

After the initial shock, I realized I had control of my thoughts and feelings. The diagnosis was also a wake-up call. I climbed mountains in my 40s, sailed the San Juan Islands, explored the reefs in Hawaii, hiked hundreds of miles of trails in the mountains and lowlands, and became a licensed pilot. I could go on, but all this happened after the symptoms appeared.

The redeeming feature of Kennedy’s Disease is it progresses slowly. I still have good days and bad days, just as I did before KD. I still become frustrated and resentful at times. I still lash out at others when it is meant for me. And, what is interesting about this is that these events would also happen if I did not have KD. It is called ‘LIFE’. I experience, adjust and adapt. But, I continue to live the best life possible.

Life is a journey of discovery. And, as with any person’s life, there are hills and valleys, as well as rivers and deserts, that have to be traversed. There will be good times and not so good times, but each will be its own learning experience. That is why this blog is called, “Living with Kennedy’s Disease… until there is a cure.

Photo: https://weheartit.com/entry/42876870

Mouth and tongue fasciculations in Kennedy’s disease

SpringLink has a video of fasciculations of the tongue and mouth.

“…Abstract - We report the case of a 54-year-old right-handed man who presented with a 2-year history of progressive upper-limb weakness with mild dysarthria and prominent involuntary perioral abnormal movements that were characterized as fasciculations…”

For those who have never experienced fasciculations, it is the strangest sensation and it is uncontrollable. The ones running up and down my back, for example, feel like I have an alien in my body trying to escape. They use to bother me. Since I have lived with them for over half of my life, I now consider them a part of the family. J

Gene Replacement Therapy for Spinal Muscular Atrophy Type 1

Even though this is not Kennedy’s Disease (SBMA) related, the gene replacement therapy mentioned in the two articles shows another major step forward in this type of treatment. If they can accomplish these type results for SMA1, the potential treatment for Spinal Bulbar Muscular Atrophy is a little closer.

The Gene Therapy Animation is short, but nicely explained in this YouTube video. 

Note: Information for parents of children with SMA1 who want to learn more about study participation can visit: studysmanow.com.

The well-written article explaining Dr. Mendell’s research in Nationwide Children’s does a good job of explaining the treatment and where they are in the process.

Phase 1 Replacement Therapy Article

“…SMA1 is a progressive, childhood, neuromuscular disease caused by a mutation in a single gene. Children with SMA1 fail to meet motor milestones and typically die or require permanent mechanical ventilation by 2 years of age. The phase 1 clinical trial is the first to test the functional replacement of the mutated gene responsible for SMA1.

A one-time intravenous injection of modified adeno-associated virus serotype 9 (AAV9) delivered the SMN gene to 15 patients. Three patients received a low dose, while 12 patients received a high dose. In the Phase 1 trial, patients in the high dose group demonstrated improvement in motor function and they had a decreased need for supportive care compared to the natural history of the disease.

Specifically, at the end of the study period, all 15 patients appeared to have a favorable safety profile and to be generally well tolerated. Of the 12 patients treated with the high dose, 92 percent of patients have achieved head control, 75 percent of patients can roll over and 92 percent of patients can sit with assistance. Seventy-five percent of these patients are now sitting for 30 seconds or longer. Two patients can crawl, pull to stand and stand and walk independently.

According to natural history of the disease, patients require nutritional and respiratory support by 12 months of age, and are not able to swallow or speak effectively. Of the patients who received the high dose in study, 11 patients are able to speak, 11 patients are fed orally and seven do not require bi-level positive airway pressure as of the data cut-off (August 7, 2017)…”